This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The hypothesis to be tested is that chemokine-induced redirected trafficking of mucosal-homing immune cells to the systemic compartment leads to mucosal cellular and humoral immune responses without mucosal immunization. Mucosal-expressed chemokines CTACK, MEC, TECK will be tested for their individual potential to enhance the mucosal immune response of a systemically delivered SIV DNA vaccine. The vaccination approach will be intramuscular co-immunization of macaques with an optimized SIVgag/pol/env plus each plasmid form of the mucosal-expressed chemokines. Specific aim 1 is in progress to measure antigen specific cell mediated immune responses systemically (PBMCs) as well as in mucosa following intramuscular plasmid co-immunization of rhesus macaques with 3 individual chemokines, CTACK, MEC, TECK, plus pSIVgag/pol/env. Three groups of 4 rhesus macaques were immunized by subcutaneous electroporation with plasmids containing the 3 mucosal chemokines plus SIVgag/pol/env. Blood, lymph nodes and vaginal and intestinal mucosal tissue was collected and tested for multi-functional T cell immune responses to SIV gag and pol. Animals responded to each of the chemokine adjuvanted vaccines. The strongest responses, compared to controls receiving SIVgag/pol/env alone, were vaginal tissue. This is consistent with the hypothesis that chemokines may redirect trafficking of mucosal-homing immune cells.